There are some unique aspects to providing trauma-informed reproductive care to transgender and nonbinary people, who are affected by minority stress, stigma, and particular forms of trauma; we review the evidence and suggest strategies for the provision of trauma-informed reproductive care to gender minorities.

In mammals, various molecules are involved in biochemical interaction between conceptus and endometrium for pregnancy recognition and establishment. In ruminants, interferon tau (IFNT) is the pregnancy recognition factor; however, IFNT alone does not explain corpus luteum maintenance. Although data on factors expressed during implantation has been accumulated, we hypothesized that the conceptus produces additional, uncharacterized molecules during the period of conceptus attachment. This study aimed to identify new conceptus secretory proteins involved in the biochemical interaction between conceptus and endometrium in sheep. We analyzed RNA-sequence data of ovine conceptuses from pregnant animals on days 12, 14, 15, 16, 17, 19, 20, and 21. To identify novel secretory proteins, we focused on highly expressed but uncharacterized genes and performed in silico protein function analysis, identifying genes encoding phospholipase inhibitory proteins expressed on days 14 and 15. Recombinant proteins from these genes were produced and the effects on cultured bovine endometrial epithelial cells (EECs) and stromal cells (STRs) were analyzed by RNA-sequence analysis. Differentially expressed gene (DEG) analysis demonstrated that the recombinant protein treatment upregulated 31 genes and downregulated 4 genes in EECs; is also upregulated 398 genes and downregulated 66 genes in STRs, including implantation-related genes such as ISG15, OAS1X, OAS1Y, PARP9, PARP14, MX1 and PTGS2. Gene set enrichment analysis revealed that DEGs were enriched in several implantation-related pathways, including ISG15 antivirus mechanisms. These results suggest that, in addition to numerous characterized molecules, phospholipase inhibitory protein is a new candidate molecule in enabling biochemical communication between the conceptus and endometrium.

Granulosa cells in the ovaries of livestock are crucial for secreting steroid hormones that regulate follicular development, with lipid synthesis and metabolism playing key roles in this process. The molecular mechanisms behind steroid hormone secretion regulated by fatty acid metabolism in goat granulosa cells have been unclear. Our previous transcriptome analysis of Yunshang black goat ovaries revealed that miR-novel-216, which had lower expression in high-fertility goats, might regulate granulosa cell function. We further investigated the role of miR-novel-216 by isolating and culturing goat granulosa cells in vitro, and found that it inhibits cell proliferation, lipid accumulation, and progesterone synthesis in goat granulosa cells. The qTar and miRanda analyses predicted TPD52 as a target of miR-novel-216, confirmed by dual luciferase and transfection assays. Previous studies have shown that progesterone synthesis in granulosa cells is closely related to free fatty acid composition. We investigated the effect of in vitro construction of TPD52 overexpression and interference plasmids on the free fatty acid content of goat granulosa cells using mass spectrometry sequencing. The results showed that overexpression of TPD52 in goat granulosa cells significantly increased free fatty acid content and promoted granulosa cell proliferation, lipid accumulation, and progesterone synthesis, whereas the opposite was true for inhibition of TPD52. It was shown that miR-novel-216 affects granulosa cell proliferation and free fatty acid levels by regulating the expression of TPD52, which increases reproductive hormone secretion and promotes polytocous trait in goats. This provides a foundation for developing breeding strategies to improve goat fertility.

Spatial genome organization refers to the conformation of the chromosomes and their relative positioning within the nucleus. In mammals, fertilization entails intense chromatin remodeling of parental genomes, as well as large-scale structural changes in nuclear organization of the newly formed zygote over the first two cell cleavages. The molecular characteristics, mechanisms and functionality of spatial genome organization during the early steps of development in mouse have been extensively studied and will be presented in this review, with a specific focus on heterochromatin. Concomitantly to maturation of genomic architecture, the embryonic genome activation occurs in transient waves of transcription. Here, we will also discuss the putative link between heterochromatin organization and the regulation of genome expression.

Metabolic syndrome (MS) is increasingly associated with impaired reproductive health. This study aimed to assess the endometrial characteristics and reproductive outcomes of a female MS mouse model and evaluate metformin's therapeutic effects. Twenty-one-day-old female C57BL/6 mice were randomly divided into a high-fat diet group (N = 50) and a control group (N = 30) that received standard chow. After 11 weeks, a subset of HF mice (N = 25) was given oral metformin at 300 mg/kg/day, while the other ones continued on HF diet. After 15 weeks, mice were either sacrificed during estrus or mated and euthanized on day 7.5 of pregnancy (N = 15 per group). The estrous cycle, progesterone and estradiol levels, uterine morphology, endometrial cell proliferation, reproductive performance, and metformin's treatment effects were assessed. Mice on a high-fat diet developed MS, which was characterized by moderate glycemic dysregulation, increased cholesterol, insulin resistance, and central obesity. Experimental MS caused estrous cycle disruptions and increased serum progesterone levels, which were normalized by metformin. MS also affected endometrial histology, producing hyperplasia and altering cell proliferation, while metformin restored normal endometrial architecture by inhibiting cell proliferation. Additionally, MS impaired reproductive success by delaying coitus and reducing the ratio of implantation sites to corpora lutea, both of which were rectified by metformin. In conclusion, MS adversely affects reproductive function, but metformin offers improvement. Our findings highlight the need for further research on the impact of MS on reproduction and the exploration of treatments to enhance reproductive health in women with MS.

Daily exposure to a mixture of phthalates is unavoidable in humans and poses a risk to reproductive health because they are known endocrine-disrupting chemicals. Specific to female reproductive health, the literature has linked phthalate exposure to impairments in ovarian function, uterine function, pregnancy outcomes, and endocrine signaling in the hypothalamus-pituitary-ovarian axis. However, limitations to these studies are that they primarily focus on single phthalate exposures in animal models. Thus, the effects of real life exposures to mixtures of phthalates and the clinical and translational impacts on reproductive function in women are largely unknown. This review summarizes recent literature specifically investigating associations between phthalate mixture exposures and clinical reproductive outcomes and reproductive disease states in women. Because these studies are scarce, they are supplemented with literature utilizing single phthalate analyses in women and mechanistic basic science studies using phthalate mixture exposures. Main findings from the literature suggest that elevated phthalate exposure is associated with altered menstrual cyclicity, altered pubertal timing, disrupted ovarian folliculogenesis and steroidogenesis, ovarian disorders including primary ovarian insufficiency and polycystic ovary syndrome, uterine disorders including endometriosis and leiomyomas, poor in vitro fertilization outcomes, and poor pregnancy outcomes. There is an urgent need to better incorporate phthalate mixtures in epidemiology (mixture analyses) and basic science (direct exposures) study designs. Further, as exposure to multiple phthalates is ubiquitous, elucidating the mechanism of phthalate mixture toxicities is paramount for improving women's reproductive health.

The morphological diversity and functional role of the organs of the female reproductive system across tetrapods (limbed vertebrates) are relatively poorly understood. Though some features are morphologically similar, species-specific modification makes comparisons between species and inference about evolutionary origins challenging. In combination with the study of morphological changes, studying differences in gene expression in the adult reproductive system in diverse species can clarify the function of each organ. Here, we use the brown anole, Anolis sagrei, to study gene expression differences within the reproductive tract of the adult female. We generated gene expression profiles of four biological replicates of the three regions of the female reproductive tract, the infundibulum, glandular uterus, and nonglandular uterus by RNA-sequencing. We aligned read to the recently published Anolis sagrei genome and identified significantly differentially expressed genes between the regions using DEseq2. Each organ expressed approximately 14600 genes and comparison of gene expression profiles between organs revealed between 367-883 differentially expressed genes. We identify shared and region-specific transcriptional signatures for the three regions and compare gene expression in the brown anole reproductive tract to known gene expression patterns in other tetrapods. We find that genes in the Hox cluster have an anterior-posterior, colinear expression pattern as has been described in mammals. We also define a secretome for the glandular uterus. These data provide fundamental information for functional studies of the reproductive tract organs in the brown anole as well as an important phylogenetic anchor for comparative study of the evolution of the female reproductive tract.

Extensive research indicates that fertilization outcomes are shaped by individual female and male traits that reflect their intrinsic quality. Yet, surprisingly little is known about the influence of interactions between the sexes and their adaptive significance in either externally or internally fertilizing species. Here, we review empirical evidence on how female-male interactions influence each stage of the fertilization process, including sperm transfer, transport, storage, chemoattraction and fertilization. We also address the challenges of examining female-male interaction effects within a realistic biological context and why research in this area lags behind understanding the role of individual sex-specific traits. While relatively little data are currently available to address interactive effects between the sexes and their impact on the fertilization process, what is presently known suggests these effects are likely to be more common across the animal tree of life than appreciated. Future research will help to not only identify these interactions, but their understanding can also help to explain the maintenance of genetic variation and inform applied studies of fertility.

Bisphenol-A (BPA), an environmental endocrine disruptor (EED), is used widely in the manufacturing of various plastics. While BPA can have detrimental effects on fertility and reproductive health, the effects of BPA on early ovarian differentiation in mammals remains unclear. Marsupials have undifferentiated gonads at birth, so this study investigated the gross morphology, protein localisation of FOXL2 and FST and the expression profile of key ovarian differentiating genes FOXL2, WNT4,FST, ESR1 and ESR2 every 2 days from the day of birth to day 10 post partum (pp) in the marsupial tammar wallaby. A second group of newborn female pouch young were treated with 50 µg/kg of BPA daily from day 0-10 pp and the morphology and gene expression were examined at day 10 pp. Ovigerous cords in tammar ovaries were first formed between days 2 to 4 pp. FOXL2 localisation became nuclear by day 4 in pre-granulosa cells. FST was initially in the cytoplasm of pre-granulosa cells at day 2 pp, but was then secreted into the extracellular matrix in ovaries by day 10 pp. FOXL2, FST, ESR1 and ESR2 mRNA were upregulated in ovaries around day 2-4 pp, indicating that ovarian differentiation in the tammar begins from day 2-4 pp. Interestingly, BPA treatment from day 0-10 pp blocked the morphological differentiation of the cortex and medulla as well as ovigerous cord formation and downregulated the expression of FST and FOXL2 at day 10 pp.

This review article summarizes the experimental findings in rodents published between 2014 and 2024 concerning phthalates exposure and reproductive outcomes. Rodents were chosen for this review since most studies that have developmental aspects in different phases of exposure and that address more in-depth reproductive mechanisms have been carried out in mice and rats. The evidence of adverse effects of phthalates on fetal development and human and animal reproduction is extensive, with impacts ranging from gene expression to physiological alterations. Despite the large volume of scientific papers pointing out the harmful effects of exposure to phthalates, isolated or in mixtures, at different developmental periods, most of them are associated with the maternal exposure and long-term effects in the offspring. Regular vegetables, fruits, fish, dairy products, and whole grains intake rich in bioactive compounds can mitigate the adverse effects of EDCs in humans and animals at different developmental periods. Various food bioactive compounds (FBCs) such as genistein, resveratrol, lycopene, vitamin E, curcumin, selenium, and plant secondary metabolites (PSMs) present antioxidant, anti-inflammatory, anti-tumor, and other biological properties with the potential to reduce of deleterious effects of phthalates on the reproductive tract. In this review, we aimed to summarize the main studies produced in the last decade about phthalate exposure and reproductive disorders in males and females (at different developmental critical windows). Additionally, we proposed some FBCs and PSMs that could attenuate the main adverse effects caused by phthalate exposure on male reproduction since there is a lack of studies with females.

Endometrial stromal cell decidualization is required for pregnancy success. Although this process is integral to fertility, many of the intricate molecular mechanisms contributing to decidualization remain undefined. One pathway that has been implicated in endometrial stromal cell decidualization in humans in vitro, is the Hippo signaling pathway. Two previously conducted studies showed that the effectors of the Hippo signaling pathway, YAP1 and WWTR1, are required for decidualization of primary endometrial stromal cells in vitro. To investigate the in vivo role of YAP1 and WWTR1 in decidualization and pregnancy initiation, we generated Progesterone receptor Cre mediated mutation of a combination of Yap1 and Wwtr1 alleles. Female Yap1 and Wwtr1 triple allele mutants exhibited subfertility, a compromised decidualization response, decreased endometrial receptivity, delayed embryonic development, and a unique transcriptional profile at 7.5 days post coitus. Bulk mRNA sequencing revealed aberrant maternal remodeling evidenced by significant alterations in extracellular matrix encoding genes at 7.5 days post-coitus in mutant dams and enrichment for terms associated with fertility-compromising diseases like pre-eclampsia and endometriosis. In addition, differentially expressed genes overlapped directionally with Estrogen receptor and Epidermal growth factor receptor regulated genes as identified by microarray. Our results indicate that Yap1 and Wwtr1 are necessary for successful mammalian pregnancy initiation.

Top-down proteomics was employed to construct proteoform atlas of sperm and seminal plasma (SP) from bulls with low (LF) and high (HF) semen freezability. Sperm and seminal proteins were fractionated by tandem size exclusion chromatography (< 30 kDa) and analyzed by reversed-phase liquid chromatography-tandem mass spectrometry. This approach enabled the identification of 299 SP (from 46 families) and 267 sperm proteoforms (from 139 families). Seventy proteoforms belonging to beta-defensin 10, c-type natriuretic peptide (NPPC), caltrin, seminal ribonuclease, osteopontin, and binder of sperm protein (BSP) 3 families were unique to HF bulls' SP. LF seminal proteins had unique 77 proteoforms, including caltrin, NPPC, osteopontin, BSP3, serpin family A member 5, and β-NGF families. Proteoform families of SP in HF and LF bulls were related to Ca2+ uptake, capacitation, acrosome reaction, sperm protection, fertilization and proteolytic processes. Thirty-three proteoforms of NPPC, caltrin, and cylicin-2 families were upregulated in HF sperm. Twenty-two proteoforms of caltrin, cylicin-2, ATP synthases, and malate dehydrogenase families were among those upregulated in LF sperm. Truncated and acetylated histone H2A and non-truncated and acetylated c-Myc binding protein were prevalent in LF sperm. Cylicin-2 proteoforms were observed in HF and LF sperm, and truncated glyceraldehyde-3-phosphate dehydrogenases, only in HF sperm. In silico analyses indicated the enrichment of mitochondrial metabolic pathways in HF sperm, including fatty acid metabolism and TCA cycle. Our study brings an unprecedented description of the bovine SP and sperm proteoforms. Post-translational processing appears to define the bio-properties of semen proteins and their associations with sperm cryoresistance.

We recently developed a three-dimensional (3-D) ovarian tissue culture system supported by bacterial-derived dextran hydrogel. Arg-Gly-Asp (RGD) is an extracellular matrix (ECM)-derived triple peptide. Immature ovarian tissues cultured in RGD-modified dextran hydrogel significantly promoted antral follicle growth and oocyte quality compared with those cultured in dextran hydrogel alone. In this study, we examined the mechanism of follicle growth stimulated by RGD treatment in the 3-D system. First, we detected that direct contact between RGD-modified dextran hydrogel and ovarian interstitial cells is necessary to promote antral follicle growth. Therefore, we hypothesized that RGD stimulates antral follicle growth through RGD-binding integrin receptors expressed in the interstitial cell mass. Using qPCR and immunochemical staining, we identified that integrins ⍺vβ3 and ⍺v5 are predominantly expressed in the ovarian interstitial compartment. To assess the effect of RGD-integrin interaction on follicle growth, ovarian tissues were cultured with Cilengitide (Ci), an inhibitor specific for ⍺vβ3 and ⍺vβ5. Ci treatment suppressed RGD-induced follicle growth and oocyte quality in a dose-dependent manner. When the interstitial cell aggregates were cultured with RGD, cell migration and theca-related gene expression were significantly upregulated. Ci treatment dramatically suppressed these RGD-induced activities. In co-culturing interstitial aggregate and secondary follicles with RGD, migrating cells formed outermost cell layers around the follicles, like theca layers, which were totally blocked by Ci treatment. In conclusion, our results suggest that RGD stimulates theca cell differentiation in the ovarian interstitial cells through integrins ⍺vβ3 and ⍺v5 to promote antral follicle growth in our 3-D system.

Years of growing research demonstrate that transgender and gender diverse (TGD) people desire fertility counseling and family building, however social and medical factors can impact future fertility options. Fortunately, TGD individuals have many viable options for family building using their own gametes and/or reproductive organs. However, the nuanced ways in which different gender affirming treatments affect reproduction, the interplay with non-treatment related infertility factors, and mitigation of likely dysphoria triggers are all critical to actual utilization. This review focuses on fertility treatment and preservation options for TGD patients and highlights these influential social and medical factors. Fertility treatments may be associated with worsening gender dysphoria in TGD people, and an affirming clinical environment and conscientious provider approach is paramount to treatment success. However, reducing gender dysphoria can also require specific changes to medically assisted reproduction and sperm collection protocols, some which carry the potential for diminished outcomes or unknown effects. Adolescents undergoing fertility preservation treatments may need more support or additional protocol modifications, and outcomes may be poorer in this age group compared with adults. Testicular and ovarian tissue cryopreservation may present a fertility preservation option for prepubertal TGD children, however in-vitro gamete maturation remains experimental in this situation.

The significant role of C-type natriuretic peptide (CNP) and its receptor 2 (NPR2) in regulating oocyte meiotic maturation and facilitating communication between oocytes and surrounding cumulus cells has been well-documented in various mammalian species, including mice, cattle and swine. However, further investigation is needed to ascertain whether natriuretic peptide receptors (NPRs) are involved in regulating other essential ovarian functions. Hence, this study aimed to explore the potential involvement of NPRs in the regulation of cumulus expansion and oocyte meiotic maturation in bovine cumulus-oocyte complexes (COCs). The findings revealed that NPR3 mRNA abundance was downregulated by FSH and LH in cumulus cells of bovine COCs during in vitro maturation (IVM), while NPR2 mRNA levels were not affected by gonadotropins. Inhibition of the epidermal growth factor receptor (EGFR) during IVM of COCs prevented the NPR3 mRNA downregulation induced by gonadotropins in cumulus cells. Additionally, treatment of COCs during IVM with an NPR3 agonist (cANP4-23) inhibited cumulus expansion induced by gonadotropins. This inhibitory effect was further intensified when COCs were co-treated with cANP4-23 and CNP. These findings provide robust evidence indicating that normal cumulus expansion in bovine COCs involves an inhibitory effect of gonadotropins on NPR3 mRNA expression, which is mediated via EGFR signaling. The study also provides evidence that CNP and NPR3 interact synergistically to regulate cumulus expansion in response to gonadotropins.

Particulate matter (PM) air pollution consists of liquid and solid particles, which are categorized by size as less than 10 (PM10) μm, 2.5 (PM2.5) μm, or 0.1μm (PM0.1 or ultrafine) in aerodynamic diameter and which vary in composition depending on the sources. PM exposure is ubiquitous and has been associated with many adverse health effects. This narrative review focuses on epidemiological and experimental studies that investigated the effects of PM exposure on female and male reproduction and on pregnancy.μ.

Ovulation is the process by which a fertilizable oocyte is extruded from the interior of the follicle. Herein, we conducted a literature survey to explore the ovulation patterns of eleven sexually reproducing species belonging to 10 animal phyla. These results indicate a large variety of ovulation patterns. Further comparative biological and evolutionary considerations of these results led us to conclude that most female animals ovulate via follicle rupture. We propose that in all animals that ovulate by follicle rupture, two cellular events may be critically involved in the process: 1) the disintegration of cell junctional systems that lead to intracellular cytoskeleton rearrangement in the follicle cells and 2) the degradation of extracellular matrix (ECM) proteins filling between follicle cells. These events may result in follicular cell deformation and increased motility, both of which are necessary for the formation of a path through which oocytes escape from the follicle. In addition to the requirement of ECM degradation for disintegrating cell junctions, intensive ECM protein degradation at the apical region of the follicle probably became increasingly important in late-evolving animals, such as vertebrates, in which a thick follicle wall containing a large abundance of ECM proteins is formed. We also considered hypothetical scenarios for the evolution of ovulation in these animals. Furthermore, this article discusses the future problems that need to be solved for a more comprehensive understanding of ovulation in the animal kingdom.

The histone variant TH2B, enriched in oocytes, sperm, and early embryos, decreases as embryos differentiate into pre-gastrula stages. Despite its presence, the role of TH2B in epigenetic reprogramming during early embryonic development remains largely under-researched. Our study employed ultra-low-input ChIP-seq (ULI-ChIP) to analyze the genome-wide distribution of TH2B in MII oocytes and early embryos. We found that TH2B is enriched in the chromatin of oocytes and 2-cell stage embryos but becomes less prevalent after the 2-cell stage. Correlation analysis revealed that the TH2B chromatin patterns in sperm and preimplantation embryos are more similar to each other than to those in MII oocytes. Gene ontology (GO) analysis of TH2B-occupied loci linked them to various developmental processes, including oogenesis, fertilization, chromatin modification, and transcription regulation. The study also identified a strong association of TH2B with specific transposable elements (TEs), particularly long terminal repeats (LTRs), which are known to regulate preimplantation development. Additionally, early embryos showed H3K9me3 marks at TH2B-bound loci. TH2B exhibited strong correlations with H2A.Z and H3.3 in the 2-cell and 8-cell stages, a positive association with H3K27Ac and H3K4me3, and a negative correlation with H3K27me3. Allelic reprogramming analysis of TH2B in embryos from C57BL/6J and DBA/2J crosses revealed differential dynamics between maternal and paternal alleles, with a notable paternal bias at the promoter in 2-cell embryos. Thus, TH2B's enrichment in early embryonic stages and its association with key regulatory regions and histone modifications underscore its importance in ZGA and subsequent developmental processes.

Exposure to endocrine-disrupting chemicals (EDCs) and pharmaceuticals during development has been linked to reproductive dysfunction, reduced semen quality, and infertility. Research indicates that EDC mixtures, which are common in the modern environment, can pose significant risks that may not be fully assessed by studying individual compound toxicity, especially at environmentally relevant doses or concentrations. Understanding the contribution of chemical mixtures to male reproductive toxicity is crucial, given the increasing reliance on pharmaceuticals and pervasiveness of anthropogenic pollution. Recent studies on EDC effects have expanded to a more diverse range of microplastics, pesticides, antimicrobials, phytoestrogens, and pharmaceuticals such as analgesics, which can collectively impact testicular function and fertility. Adverse outcomes observed across studies include reproductive tract malformations, decreased sperm count and motility, lowered testosterone, delayed-onset puberty, and possible causal effects such as oxidative stress and altered gene expression. Still, limited data exists on combinations of environmental pollutants and pharmaceuticals with ED potential at human-relevant doses. This review of the recent literature aims to synthesize the toxicological impact of low dose chemical mixtures on male reproductive health. Overall, humans are exposed to EDCs and drugs through various ways, necessitating an understanding of their concomitant effects on male reproductive health.

Brain-derived neurotrophic factor (BDNF) is a peptide widely known for its role in neurogenesis and synaptic plasticity. Its expression in non-neuronal tissues has been reported. In mammals, it is involved in ovarian development, follicle growth, oocyte maturation, and early embryonic development. In zebrafish, it was demonstrated that BDNF increases food intake and regulates metabolism. Reproduction and metabolism are tightly linked. We hypothesized that BDNF modulates reproductive hormones and reproductive functions in zebrafish. This study aimed to determine BDNF expression in the zebrafish reproductive axis and whether it modulates the reproductive endocrine milieu and oocyte biology in zebrafish. Our results show that bdnf and its receptor trkb, and BDNF-like immunoreactivity are detected in zebrafish gonads and liver cells. This suggests BDNF local production and possible actions within the gonads and liver. Intraperitoneal administration of 1, 10, or 100 ng/g bodyweight BDNF significantly (ANOVA, p<0.05) increased sgnrh/cgnrh-II, kiss1, and cyp19a1b mRNAs in the zebrafish brain; steroidogenic enzymes (star and cyp19a1a) and key receptors in the zebrafish gonads. In vitro incubation of zebrafish liver cells with BDNF significantly (ANOVA, p<0.05) increased estrogen receptor mRNAs and vitellogenin concentrations (ELISA) in the cells. BDNF (100 ng/mL) induced (ANOVA, p<0.05) oocyte maturation in vitro at 24 hours post-incubation and significantly upregulated cumulus-expansion related genes (ANOVA, p<0.05). Overall, our findings indicate a stimulatory role for BDNF in the reproductive axis of zebrafish. This provides impetus for future research on its mechanism of action and potential practical applications to enhance reproduction in aquaculture.

Placenta-associated pathologies, including early pregnancy loss (EPL) and preeclampsia (PE), share a common phenomenon of insufficient extravillous trophoblasts (EVTs) invasion. It was previously observed that down-regulated miR-486-5p expression inhibited the invasion of EVTs, and a decreased peripheral miR-486-5p was associated with EPL. However, the exact roles of miR-486-5p played in pathogenesis of EPL, as well as the molecular pathway underlying roles of miR-486-5p in EVTs invasion, remains poorly understood. In this study, a decreased miR-486-5p expression in uterine embryo implantation site at gestation day (GD) 8.5, and an increased expression of Smad2, a target of miR-486-5p, were observed in the lipopolysaccharide (LPS)-induced EPL mouse model. The invasion and viability of immortalized human EVTs line, HTR-8/SVneo, were inhibited by LPS, accompanied with a reduced miR-486-5p expression. LPS showed a promoting effect on the Smad2 expression, of which could be attenuated by miR-486-5p mimics. And the down-regulated Smad2 could effectively restore the impaired invasion and viability of HTR-8/SVneo cells caused by LPS or miR-486-5p inhibitor. Furthermore, LPS could promote the TNFα production in HTR-8/SVneo cells, whereas both of siSmad and miR-486-5p mimics could reverse such an effect. By analyzing the human decidua single-cell RNA sequencing and transcriptome datasets derived from the Gene Expression Omnibus, it was found that, compared to control early pregnant women, the Smad2 expression was significantly increased in recurrent miscarriages (RM) patients. Collectively, these data suggested that, decreased miR-486-5p expression might lead to EPL at least partially by inhibiting invasion and/or promoting TNFα production of EVTs via targeting Smad2.